4-hydroxy-steroids and process of their preparation



3,068,225 4-HYDROXY-STEROIDS AND PROCESS OF THEIR PREPARATIQN Bruno Camerino, Milan, Bianca Patelli, Stradella, and

Roberto Seiairy, Milan, Italy, assignors to Societa Farmaceutici Italia, Milan, Italy, a corporation of Ital No Drawing. Filed Aug 4, 1961, Ser. No. 129,255 Claims priority, application Italy Oct. 30, 1960 14 Claims. (Cl. 260-23955) Our invention relates to a process for obtaining the 4-hydroxy-steroids of the androstane series of l9-norandrostane, of l7a-hydroxy-pregnane, its 21-hydroxylated derivative, and the new products of said process.

More particularly, our invention refers to the oxidation with oxygen, in the presence of the potassium salt of a t.aliphatic alcohol, of the S-keto-M-steroids of the androstane series of 19-nor-androstane, of l7a-hydroxy-pregnane and l7a,2l-dihydroxy-pregnane to 4-hydroxy-3-keto- M' -steroids, from which the corresponding 4-hydroxy- 3-keto-A -steroids are obtained by catalytic hydrogenation of the A.

In the copending application of Camerino et 211., Serial No. 129,478, filed August 4, 1961, entitled Process for the Manufacture of 4-Hydroxy-3-Keto-A -Steroids, a process was described and claimed for obtaining the 4- hydroxy-3-keto-A -steroids (C) of the androstane series of 19-nor-androstane, of 17u-hydroxy-pregnane and 17a, Zl-dihydroxy-pregnane, from the corresponding 3-ket0- Sfl-steroids (B), which are obtained by catalytical hydrogenation of the B-keto-M-steroids (A), by reaction with oxygen in the presence of the potassium salt of a t.aliphatic alcohol. Although good yields are obtained, this process still involves the reduction of the 3-keto-A steroids (A) to the 3-keto-5B-steroids (B), which is not a quantitative reduction owing to the fact that 500 isomers are also always formed, to greater or lesser degree.

We have found, and this is an object of the present invention, that 3-keto-A -steroids (A) of the androstane series of l9-nor-androstane, l7a-hydroxy-pregnane and the latters Zl-hydroxylated derivative, when treated with oxygen in the presence of the potassium salt of a t.aliphatic alcohol, yield the corresponding 4-hydroxy-3-keto- M' -steroids (D), from which the relative 4-hydroxy-3- keto-M-steroids (C) are obtained in very good yield by the catalytic hydrogenation of A where R=H or CH;,.

Sttes Patent According to the present invention, 3-keto-A -ster0ids of the androstane series of 19-nor-androstane and of 17a;- hydroxy-pregnane; the latter having a 17,20,20,2 1-bismethylendioxy or substituted bismethylendioxy group when a dioxyacetonic chain is present in C17 or, but not necessarily, an ethylendioxy group in CZO when an acetylic chain is present in C17; are dissolved in t.aliphatic alcohols, such as t.butyl alcohol or t.amyl alcohol and reacted either with oxygen 01' air in the presence of the potassium salt of a t.aliphatic alcohol, such as potassium t.butylate or potassium t.amylate, at a temperature range between 10 and 50 C., preferably at room temperature for a period of time varying from a few hours to several days.

When the reaction is completed, the reaction mixture is neutralized with acids, such as acetic acid, or diluted inorganic acids, then diluted with water and the steroid subsequently extracted with a usual water-immiscible organic solvent. The resulting crude products, i.e. the 4- hydroxy-3-keto-A -steroids, may be isolated in pure form either by crystallization from organic solvents or by chromatography through resins such as Florisil (trademark) and successive elution and crystallization, or may be transformed into the corresponding acylates by acylation of the present primary and secondary hydroxyls with a chloride or anhydride of an acid in the presence or absence of t.amines, or converted into the corresponding 4-hydroxy-3-keto-A -ster0ids by catalytic hydrogenation of the A The protecting groups: of the side chain in 0-17, eventually present, of the 4-hydroxy-3-keto-A steroids obtained after oxidation may be removed before or after the acylation of the C4 hydroxyl or the hydrogenation of the A The present invention refers to the process for obtaining 4-hydroxy-3-keto-A -steroids from the 3-keto-A -ste roids. The process may be carried out with either atmospheric oxygen or pure oxygen. The air or oxygen may either be blown into the solution, in which case any carbon dioxide and moisture, if present, are preferably removed, or reacted on the surface as by allowing the solution of the steroid to stand in large and open containers.

The molar ratio of potassium t.alcoholate to steroid may vary from 1 to 30, preferably between 5 and 10. Yields are approximately of S0-90%.

The transformation of the 4-hydroxy-3-keto-A -steroids into 4-hydroxy-3-keto-A -steroids is carried out by hydrogenation in the presence of catalysts, such as 510% palladium over charcoal or platinum dioxide to absorb one mole of hydrogen, at room temperature and pressure followed by purification according to the techniques stated above with respect to the A -steroids. The yields of the selective hydrogenation of the double bond in 6-7 position are practically quantitative.

4-hydroxy-testosterone (II), 4,1113 dihydroxy 17amethyl-testosterone (IV), 4-hydroxy-17a-methy1-testosterone (VI), 4-hydroxy-l9-nortestosterone (VIII), 4-hydroxy-cortisone (X), 4-hydroxy-hydrocortisone (XH), 4, 17a-dihydroxy-progesterone (XIV), and the corresponding A analogues and the derivatives thereof were prepared according to the present process.

As far as it is known, among the group of the 4-hydroxy-3-keto-A -steroids only the A and the A -4-hydroxy-(or 4-acyloxy)-l7a-methyl-testosterones, showing a high anabolic activity when administered orally, have been described (Camerino et a1. application Serial No. 79,960, filed January 3, 1961).

The 4-hydroxy-3-keto-A -steroids obtained according to the present process are useful because the therapeutically useful corresponding 4-hydroxy-3-keto-A -steroids are obtained from them and because they also possess interesting hormonal properties.

1 g. of testosterone is dissolved in 40 cc. of t.butyl alcohol. A solution of potassium t.butylate, which is prepared by dissolving 450 mg. of potassium in cc.

of t.butyl alcohol, is added to the testosterone solution. The reaction mixture is left standing at room temperature for 24 hrs. and is occasionally stirred. Thereafter 1 cc.

of CH3COOH is added, the solution is diluted with ethyl acetate, then poured into a saturated aqueous solution of sodium chloride. The organic layer is separated, Washed to neutrality with H O, dried on Na SO and evaporated to dryness. The residue is dissolved in benzene, filtered and chromatographically separated through g. of Florisil. By using benzene and mixtures of benzene-ether 9:1

' (in volume), a product is eluted which is slightly soluble in ether and crystallizes from methanol and afterward from acetone, melting at 210-215 C.; A at 318 m =21,200; and red-violet coloration with FeCl This is A -androstadiene-4, l7fi-diol-3-one (I) EXAMPLE 1A 4,17-Diacetate of A -Androstadiene-4J7 8-Di0l-3-One mg. of A -androstadiene-4,17fi-diol3-one (I) are dissolved in 0.2 cc. of pyridine and acetylated with 0.06 cc. of acetic anhydride for 1 hour on a boiling waterbath. Dilution with H O causes the product to precipitate which is filtered off, washed with water, dried and recrystallized from acetone-petroleum ether.

This is the 4,17-diacetate of I melting at ISO-153 G; A at 288 m e=22,600.

EXAMPLE 2 4-Hydr0xy-Test0ster0ne (II) From I 25 mg. of A -androstadiene-4,17B-diol-3-one are dissolved in 2 cc. of anhydrous ethyl alcohol and hydrogenated at room temperature and pressure, in the presence of 2 mg. of PtO the reaction is stopped when one mole of hydrogen is absorbed. The catalyst is filtered off and the solution evaporated to dryness; the semicrystalline residue melts at 190-215" C. and has an UV. absorption "maximum at 278 mu.

By crystallizing from acetonepetroleum ether, II is obtained in pure form, M.P.=221- 223 C.

' EXAMPLE 3 1 7a-M ethyl-A -A ndrostadiene-4,1 1 5,1 7 5- Triol-3-One (III) A solution of potassium t.butylate, prepared from 450 mg. of potassium and 15 cc. of t.butyl alcohol, is added to 1 g. of 1lfi-hydroxy-l7oa-methyl-testosterone dissolved in cc. of t.butyl alcohol. The reaction. is kept at room 9 temperature for 24 hrs. with occasional stirring.

'of-CH COOH is then added to the solution, which is vsequentially diluted'with ethyl acetate, and poured into a saturated aqueous solution of sodium chloride. The

\ organic layer is'separated, washed with 5% aqueous solution of Na CO and with water to neutrality. The prodnet is dried on anhydrous Na SO and evaporated to dryness. The residue is .dissolved in benzene, filtered and chromatographically separated through 30 g. of Florisil.

Elution from the column with benzene and with mixtures of benzene-ether 9:1 and 8:2 (in volume) produces a product which is slightly soluble in ether and melts at 190-200 C. when recrystallized from' acetone-ether; X at 318mg; The product gives a red-violet coloration with FeC1 This is 17a-methyl-A -androstadiene- 4, 1 1/3, 17,8-triol-3-one (III) anhydrous Na SO and evaporated to dryness.

4 EXAMPLE 4 4,11,B-Dihydwxy-I7a-Methyl-Testosterone (IV) From III The hydrogenation of 17a-methyl A -androstadiene- '4,11/8,17/3,triol-3-one. (III), in the presence of 10% palladium over charcoal, at room temperature and pressure with absorption of one mole of hydrogen, yields 4,11 3- dihydroxy-l7a-methyl-testosterone (IV), melting at 184- 186 C.

EXAMPLE 5 17a-Methyl-A -Androstadiene-4J7,6-Diol-3-One (V) 5 g. of 170t-IHCII1Y1-IGSIOSI6I'OII6 dissolved in 200 cc. of t.butyl alcohol and cc. of t.butyl alcohol, wherein 2.25 g. of potassium were dissolved, are left together in an open flask at room temperature overnight. The mixture is acidified with acetic acid and extracted with ethyl acetate. The extract is Washed with a saturated NaCl solution, then with 10% sodium bicarbonate solution and finally with water to neutrality. After evaporation of the solvent, a residue of 4.900 g. is obtained which is chromatographically purified through Florisil. From the eluted fractions with benzene and ether-benzene in 9:1 volume ratio, 17m-methyl-A -androstadiene-4,17/3-diol-3- one (V), which melts at 209211 C., crystallizes out.

EXAMPLE 6 4-Hydr0xy-17a-Methyl-Test0ster0ne (VI) From V 1 g. of 17a-methyl-A -androstadiene-4,175-diol-3-one (V) are dissolved in 50 cc. of anhydrous ethyl alcohol and hydrogenated at room temperature and pressure in the presence of 25 mg. of 10% palladium over charcoal. After absorption of one mole of hydrogen, the hydrogenation speed decreases remarkably; then the reaction is stopped, the catalyst filtered off, and. the solution evaporated to dryness under vacuum. The crude product has an UV. absorption maximum at 278 me. The crude product is dissolved in chloroform, then passed through a column of Florisil with sequential elution with'chloroform; the chlorotormic eluates are evaporated to dryness and the crystalline residue is pulped with a mixture of ether and petroleum ether. The product is thereafter filtered and dried to obtain the 4-hydroxy-17wmethyltestosterone (VI) melting at -172 C.

EXAMPLE 7 19-N0r-A -AnJr0stadienc-4J7fi-Diol-3-One (VII) 3 g. of l9-nor-testosterone are dissolved in 120 cc. of t. 'butyl alcohol. A solution of potassium t.butylate prepared from 1350 mg. of potassium and 45 cc. of t.butyl alcohol is added to the 19-nor-testoste1'one solution. The mixture is permitted to react for 15 hrs. at room temperature with occasional stirring. The product is acidified with 3 cc. of acetic acid, diluted with ethyl acetate and poured into an aqueous saturated NaCl solution. The extract is then separated, washed with 5% solution of Na CO and with H O to neutrality. It is then dried on The residue is dissolved in benzene and chromatographically purified through 50 g. of Florisil. Eluting with mixtures of benzene-ether 9:1 (in volume), a product is obtained which when recrystallized from acetone-ether melts at -200 C.; A at 316 mu; e=18,600. The product gives a red-violet coloration with FeCl This is the 19- nor-A -androstadiene-4,17,8-diol-3-one (VII) I EXAMPLE 8 4-Hydroxy-19-Nor-Testosterone (VIII) From VII By hydrogenation of the l9-nor-A -androstadiene-4, l7fl-diol-3-one (VII), in the presence of 10% palladium over charcoal and at room temperature to absorption of one mole of hydrogen, the 4-hydroxy-l9-nor-testtosterone (VIII) is obtained (M.P. 187189 C.).

5 EXAMPLE 9 A -Pregnadiene-4-0l-3J I -Dine-1 7 oc,2 0,20,21 J? ismethylene Dioxy (IX) 15 cc. of t.butyl alcohol containing 0.450 g. of potassium are added to 1 g. of cortisone-17,20,20,21-bismethylenedioxy suspended in 50 cc. of t.butyl alcohol. The product is left at room temperature overnight and dissolves during reaction. When acidification with acetic acid is over, the solution is extracted with ethyl acetate. The extract is then washed with a saturated salt (NaCl) solution, then with a solution of 10% sodium bicarbonate and finally with water to neutrality. After distillation of the solvent, the residue (0.900 g.) is crystallized from methylene chloride-methanol. A -pregnadiene-4-ol-3, 11 dione 17ot,20,20,2l-bismethylenedioxy is obtained (M.P.=3l03l5 C.; A at 318 m e=20,300).

EXAMPLE 9A 4-A cetate of A -Pregnadiene-4Ol-3,I1-Di0ne-]7oc,20, 20,21 -Bismethylenedi0xy 250 mg. of the preceding product dissolved in 3 cc. of pyridine are acetylated at room temperature for 12 hrs. with 0.75 cc. of acetic anhydride. The corresponding 4- acetate melting at 195197 C is isolated in the known manner.

EXAMPLE 9B A -Pregnaaiene-4J 7 (1,21 -Triol-3,1 J ,ZO-Trione (IX) 200 mg. of A -pregnadie11e-4-ol-3,1l-dione-l7a,20,20, 2l-bisrnethylenedioxy are heated for two hours at 100 C. in 20 cc. of 60% aqueous formic acid to obtain at the end A -pregnadiene4,17a,2l-tri0l-3,l1,20-trione (IX) melting at 203-205 C.; A at 318 III/.0; e=22,500.

EXAMPLE 10 4-Hya'r0xy-C0rtis0ne (X) From IX By hydrogenation of the A -pregnadienel,17a,21-triol- 3,11,20-trione (IX), in the presence of 10% palladium over charcoal, at room temperature and pressure to absorption of one mole of hydrogen, 4-hydroXy-cortisone (X) is obtained (M.P.=215218 C.).

EXAMPLE 11 A -Pregnadiene-4-1JB-Diol-S-One-J 7a,20,20,21 -Eismelhylene-dioxy g. of hydrocortisone 17,0,20,21-bismethylene-dioxy, when treated with potassium in t.butyl alcohol as described in Example 9, yield 4.5 g. of crude product which are crystallized from methylene chloride-methanol. This is the A -pregnadieneifl1,8-diol-3-one-17a,20,20,21-bismethylenedioxy melting at 288-290 C.; A at 318 mu; e=20,670.

EXAMPLE 11A A -Pregnadiene-4,115,1 7a,21-Tetr0l-3,20-Di0ne (X1) 250 mg. of the preceding product are heated for two hours at 100 C. in 20 cc. of 60% aqueous formic acid and yield A -pregnadiene-4,11,8,17a,21-tetrol-3,2O-dione (XI) melting at 2l8-220 C.; A at 318 my; e=22,800.

EXAMPLE 12 4-Hydr0xy-Hydrocortisone(XII) From XI The 4-hydroxy-hydrocortisone (XII) is obtained by hydrogenation of A -pregnadiene-4,11,3,17m,21-tetro1-3, 20-dione (XI), in the presence of PtO and at room temperature and pressure by absorption of one mole of hydrogen (M.P.=205-208 C.).

EXAMPLE 13 A -Pregrzadiene-4J 7a-Di0l-3-One-20-Ethylene-Glycol- Ketal-4-Acetate 5 g. of 17a-hydroxy-progesterone-20-ethylene-glycol EXAMPLE 13A d -Pregnaziiene-il 7ot-Di0l-3,20-Di0ne (XIII) 0.200 g. of the product of Example 13 are heated for 2 hours at C. in 20 cc. of 60% aqueous formic acid to yield n -pregnadiene-4,l7a-diol-3,20-dione (XIII) melting at 263265 G; A at 318 m e=23,760.

EXAMPLE l4 4,17ct-Dihydroxy-Progesterone (XIV) From XIII By hydrogenation of the A -pregnadiene-4,l7a-diol-3, 20-dione (XIII), in the presence of 10% palladium over charcoal and at room temperature and pressure by absorption of one mole of hydrogen, 4,17a-dihydroxy-progesterone (XIV), melting at 228230 C., is obtained.

We claim:

1. A process for preparing 4-hydroxy-3-keto-A -steroids having the following formula:

iii )0 wherein X is selected from the group consisting of H H(OH) and O; R is selected from the group consisting of H and CH R" is selected from the group consisting of when R is selected from the group consisting of H and CH R is selected from the group consisting of and ( 3113 CHZOH OH2-0"OH2 O O O and O- 0 i-OR -o R CH2 When R is CH and R is selected from the group consisting of H and -COCH which comprises reacting the corresponding 3-keto-A steroid, in a t.aliphatic alcohol medium in the presence of the potassium salt of a t.aliphatic alcohol, with an oxygen-containing gas and purifying the resultant 4-hydroxy-3-keto-A -steroids.

2. A process for preparing 4-hYdi'OXY-3-kfiiO-A -Steroids having the formula of claim 1, which comprises reacting the corresponding 3-keto-A -steroids, in a t.aliphatic alcohol medium in the presence of the potassium salt of a t.aliphatic alcohol, with an oxygen-containing gas and acetylating the resultant 4-hydroxy-3-keto-A -steroids.

3. A process for preparing 4-hydroxy-3-keto-A -steroids having the formula of claim 1, which comprises reacting the corresponding 3-keto-A -steroid, in a t.aliphatic alcohol medium in the presence of the potassium salt of a t.aliphatic alcohol, with an oxygen-containing gas and catalytically hydrogenating the resultant 4-hydro-3-keto- A -steroids into the corresponding 4-hydroXy-3-keto-A accesses 7 steroid, by a catalyst selected from the group consisting of -10% palladium over charcoal and platinum dioxide.

4. A process forpreparing 4-hydroXy-3-keto-A -steroids having the formula of claim 1, which. comprises reacting the'cor'responding 3-keto-A -steroid, in a medium comprising a t.aliphatic alcohol selected from the group consisting of t.b'utyl alcohol and t.amyl alcohol, in the presence .of the potassium salt of a t.aliphatic alcohol selected from the group consisting of t.butyl alcohol and t.amyl alcohol, with air, said air being blown into the solution after removal of any carbon dioxide and moisture that may be present in said air, at a temperature between C. and 50 C., and purifying the resultant 4-hydroxy- 3-ke'to-A -steroid.

5. A process for preparing 4-hydroxy-3-keto-A -steroids having the formula of claim 1, which comprises reacting the corresponding 3-keto-A -steroid, in a medium comprising a t.aliphatic alcohol selected from the group consisting of tlbutyl alcohol and t.amyl alcohol, in the presence of the potassium salt of a t.aliphatic alcohol selected from the group consisting of t.butyl alcohol and t.amyl alcohol, with air, said air being blown into the solution after removal of any carbon dioxide and moisture that may be present in said air, at a temperature between 10 C. and 50 C., acetylating the resultant 4-hYdIOXY-3- keto-A -steroid and hydrogenating in the presence of a hydrogen catalyst selected from the groupconsisting of 5l0% palladium over charcoal and platinum dioxide to produce the corresponding 4-hydroxy-3-keto-A steroid.

6. l7a-methyl-M -androstadiener,l 1,8,17B-triol-3-onc.

7. A -pregnadienet,17a,21-triol-3,11,20-trione.

8. A -pregnadiene-4-ol-3,ll-dione 17a,20,20,21 bismethylenedioxy.

9. A -pregnadiene-4-ol-3,ll-dione 17u,20,20,21 bismethylenedioxy-4-acetate.

l0. A -pregnadiene-4,11/3,17a,21-tetrol-3,20-dione.

11. A -pregnadiene-4,1 lB-diol-3-one-17u,20,20,21-bismethylenedioxy.

l2. A -pregnadiene-4,l7a-diol-3,20-dione.

13. A -pregnadiene-4,17a-diol 3 one 20 ethyleneglycol-ketal.

6 -14. 4-hydroxy-3' l eto-A -steroids having the following structural formula:

Wherein R is selected from the group consisting of H and CH R" is selected from the group consisting of 0 R i---H when R is H and X is selected from the group cons sting of 5(H)OH and O; R" is selected from the group consisting of O R la and OR l--CHa when R is CH and X is selected from the group consisting of ;8(H)OH and O; Q R" is selected from the group consisting of Inhofien et al Feb. 1; 1944' Colton Mar. 13, 1956 and UNITED STATES PATENT OFFICE CERTIFICATE or UQRECTION Patent No 3,068.225 December 11, 1962 Bruno Camerino et a1.

It is hereby certified that error appears in the above numbered patent reqdiring correction and that the said Letters Patent should read as corrected below.

In the heading to the printed specification, line 9, for "Claims priority, application Italy Oct. 30, 1960" read Claims priority, application Italy Oct, 3, 1960 Signed and sealed this 21st day of May 1963 (SEAL) Attest:

ERNEST w. SWIDER DAVID LADD Attesting Officer Commissioner of Patents 

1. A PROCESS FOR PREPARING 4-HYDROXY-3-KETO-$4,6-STEROIDS HAVING THE FOLLOWING FORMULA: 